Chymotrypsin, Urease Inhibitory and Antioxidant Activities of Malva neglecta Wallr.

Öz

Objective: In this study, determining the chymotrypsin and urease inhibitory activities as well as antioxidant activity, total phenol and flavonoid content of Malva neglecta Wallr. were aimed. Method: Between April-June 2021, methanol and water extracts were prepared from the flowering and fruiting aerial parts of the plant. Then, in vitro urease and chymotrypsin inhibitory activity were determined. ABTS, CUPRAC and DPPH methods were used to determine the antioxidant activity. Total phenol and flavonoid contents were determined using Folin Ciocalteu and aluminum chloride colorimetric methods, respectively. Results: The methanol extract showed significant chymotrypsin inhibitory activity (IC50: 67.67 µg/mL); on the other hand, both water and methanol extracts moderately inhibited the urease enzyme (36.52 and 34.38% inhibition, respectively). While the methanol extract exhibited higher antioxidant activity in DPPH and CUPRAC tests; the water extract showed higher radical scavenging activity in the ABTS test. In parallel with the results obtained in the DPPH and CUPRAC tests, flavonoid content of the methanol extract was found to be higher (42.93 mg rutin equivalent/g). Conclusion: In conclusion, Malva neglecta attracted attention with its potent chymotrypsin inhibitory activity. Also considering its urease inhibitory activity and antioxidant activity parallel to its flavonoid content, it was concluded that more comprehensive studies should needs to be done on this plant.

Anahtar Kelimeler

• Malva neglecta
• chymotrypsin
• urease
• medicinal plant

Malva neglecta Wallr. Bitkisinin Kimotripsin, Üreaz İnhibe Edici ve Antioksidan Aktivitesi

Öz

Amaç: Bu araştırmada Malva neglecta Wallr.’nın kimotripsin ve üreaz enzimlerini inhibe edici aktivitesinin tespiti ve ayrıca antioksidan aktivitesi, toplam fenol ve flavonoit içeriğinin belirlenmesi amaçlanmıştır. Yöntem: 2021 yılı Nisan-Haziran ayları arasında bitkinin çiçekli ve meyveli toprak üstü kısımlarından metanol ve su ekstresi hazırlanmış ve bu ekstrelerin in vitro üreaz ve kimotripsin inhibitör aktivitesi belirlenmiştir. Antioksidan aktivitenin tespiti için ABTS, CUPRAC ve DPPH yöntemleri kullanılmıştır. Toplam fenol ve flavonoit içerikleri ise sırasıyla Folin Ciocalteu ve aluminyum klorür kolorimetrik yöntemleri kullanılarak belirlenmiştir. Bulgular: Metanol ekstresi önemli düzeyde kimotripsin inhibe edici aktivite göstermiştir (IC50: 67.67 µg/mL); üreaz enzimi üzerinde ise hem su hem metanol ekstresi orta düzeyde inhibe edici etki göstermiştir (sırasıyla %36.52 ve 34.38 inhibisyon). DPPH ve CUPRAC testlerinde metanol ekstresi daha yüksek antioksidan aktivite sergilerken; ABTS testinde su ekstresi daha yüksek radikal süpürücü aktivite göstermiştir. DPPH ve CUPRAC testlerinde elde edilen sonuçlarla paralel olarak metanol ekstresinin flavonoit içeriğinin daha yüksek olduğu anlaşılmıştır (42.93 mg rutin eşdeğeri/g). Sonuç: Sonuç olarak M. neglecta kuvvetli kimotripsin inhibe edici aktivitesi ile dikkat çekmiştir. Üreaz inhibe edici aktivitesi ve flavanoit içeriğiyle paralel antioksidan aktivitesi de dikkate alındığında bitki üzerinde daha kapsamlı çalışmaların yapılması gerektiği sonucuna varılmıştır.

Anahtar Kelimeler

• Malva neglecta
• kimotripsin
• üreaz
• antioksidan
• tıbbi bitki

Kaynakça

1. 1. Amtul Z, ve ark. Cysteine based novel noncompetitive inhibitors of urease(s)-Distinctive inhibition susceptibility of microbial and plant ureases. Bioorg Med Chem 2006;14:6737-44.
2. 2. Yılmaz Ö, Okçu N. Helicobacter pylori ve gastrointestinal sistemle ilişkili hastalıklar. Atatürk Üniv Tıp Der 2006;38:13-7.
3. 3. Korona-Glowniak I, ve ark. The in vitro activity of essential oils against Helicobacter pylori growth and urease activity. Molecules 2020;25(3).
4. 4. Upadhyay LSB. Urease inhibitors: a review. Indian J Biotech 2012;11:381-8.
5. 5. Matczuk D, Siczek A. Effectiveness of the use of urease inhibitors in agriculture: a review. Int Agrophysics 2021;35:197-208.
6. 6. Modolo LV, ve ark. An overview on the potential of natural products as ureases inhibitors: A review. J Adv Res 2015;6:35-44.
7. 7. Shi X-Z, Zhao X-F, Wang J-X. Molecular cloning and expression analysis of chymotrypsin-like serine protease from the Chinese shrimp, Fenneropenaeus chinensis. Fish Shellfish Immunol 2008;25:589-97.
8. 8. Legerská B, Chmelová D, Ondrejovič M. TLC-Bioautography as a fast and cheap screening method for the detection of α-chymotrypsin inhibitors in crude plant extracts. J Biotech 2020;313:11-7.

9. 9. Awosika T, Aluko RE. Enzymatic pea protein hydrolysates are active trypsin and chymotrypsin inhibitors. Foods 2019;8(6):200.
10. 10. Blanco A, Blanco G. Amino Acid Metabolism. In: Blanco A, Blanco G, Editors. Medical Biochemistry. USA: Academic Press; 2017. pp: 367-99.
11. 11. Gitlin-Domagalska A, Maciejewska A, Debowski D. Bowman-Birk inhibitors: Insights into family of multifunctional proteins and peptides with potential therapeutical applications. Pharmaceuticals 2020;13:421.
12. 12. Cid‑Gallegos MS, ve ark. Protease inhibitors from plants as therapeutic agents‑ A Review. Plant Foods Human Nutr 2022;77:20-9.
13. 13. Kårlund A, ve ark. Intestinal exposure to food-derived protease inhibitors: digestion physiology- and gut health-related effects. Healthcare 2021;9:1002.
14. 14. Chen Z, ve ark. Sulforaphane is a reversible covalent inhibitor of 3‐chymotrypsin‐like protease of SARS‐CoV‐2. J Med Virol 2023;95:e28609.
15. 15. Tijjani H, ve ark. Pharmacoinformatic study of inhibitory potentials of selected flavonoids against papain-like protease and 3-chymotrypsin-like protease of SARS-CoV-2. Clin Phytosci 2022;8: https://doi.org/10.1186/s40816-022-00347-y
16. 16. Gulcin İ. Antioxidants and antioxidant methods: an updated overview. Arch Toxicol 2020;94:651-715.
17. 17. Sezik E. Türkiye'de Halk İlaçları Araştırmaları ve Önemi. 9. Bitkisel İlaç Hammaddeleri Toplantısı Bildiri kitabı; 1991; Mayıs 19-16; Eskişehir: Anadolu Üniversitesi Yayınları;1991.
18. 18. Gertsch J. How scientific is the science in ethnopharmacology? Historical Perspectives and epistemological problems. J Ethnopharmacol 2009;122:177-83.
19. 19. Agthael MAV, Eggelte T, Boxtel CJV. Artemisinin drugs in the treatment of malaria: From medicinal herb to registered medication. Trends Pharm Sci 1999; 20:199-205.
20. 20. Heinrich M, Teoh HL. Galatamine from snowdrop-the development of a modern drug against Alzheimer’s disease from local caucasian knowledge. J Ethnoparmacol 2004;92(2-3):147-62.
21. 21. Akgül A, ve ark. An ethnobotanical study in Midyat (Turkey), a city on the silk road where cultures meet. J Ethnobiol Ethnomed 2018;14(12).
22. 22. Altundağ E, M MÖ. Ethnomedicinal studies on the plant resources of east Anatolia, Turkey. Procedia Soc Behav Sci 2011;19:756-77.
23. 23. Badem M, ve ark. Biological screening of traditional medicinal plants from villages of Akkus (Ordu) in Turkey on the effects of tyrosinase. J Pharm Res Int 2018;25(6):1-10.
24. 24. Gürbüz İ, ve ark. Folk medicine in Düzce province (Turkey). Turk J Bot 2019;43(6):769-84.
25. 25. Bulut G, Tuzlacı E. An ethnobotanical study of medicinal plants in Turgutlu (Manisa-Turkey). J Ethnopharmacol 2013;149:633-47.
26. 26. Khan KM, ve ark. Synthesis and urease enzyme inhibitory effects of some dicoumarols. J Enzyme Inh Med Chem 2004;19(4):367-71.
27. 27. Cannell RJ, ve ark. Results of a large scale screen of microalgae for the production of protease inhibitors. Planta Med 1988;54(01):10-4.
28. 28. Matthaus B. Antioxidant activity of extracts obtained from residues of different oilseeds. J Agricult Food Chem 2002;50:3444-52.
29. 29. Apak R, ve ark. A novel total antioxidant capacity index for dietary polyphenols, vitamin C and E, using their cupric ion reducing capability in the presence of neocuproine: CUPRAC method. J Agricult Food Chem 2004;52:7970-81.
30. 30. Gülçin İ. Comparison of in vitro antioxidant and antiradical activities of L-tyrosine and L-Dopa. Amino Acids 2007;32(3):431-8.
31. 31. Woisky RG, Salatino A. Analysis of propolis: some parameters and procedures for chemical quality control. J Apicultl Res 1998;37:99-105.
32. 32. Singleton VL, Rossi JA. Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents. American J Enol Viticult 1965;16:144-58.
33. 33. Singleton VL, Orthofer R, Lamuela-Raventos RM. Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu Reagent. Method Enzymol 1999;299:152-78.
34. 34. Nabati F, ve ark. Large scale screening of commonly used Iranian traditional medicinal plants against urease activity. J Pharm Sci 2012;20(72).
35. 35. Biglar M, ve ark. Screening of 20 commonly used Iranian traditional medicinal plants against urease. Iran J Pharm Res 2014;13(Suppl):195-8.
36. 36. Akkol EK, ve ark. The phytochemical profile and biological activity of Malva neglecta Wallr. in surgically induced endometriosis model in rats. Molecules 2022;27:7869.
37. 37. Son NT, ve ark. Serine protease inhibitors and activators from Dalbergia tonkinensis species. J Nat Med 2020;74:257-63.
38. 38. Nile SH, ve ark. Utilization of quercetin and quercetin glycosides from onion (Allium cepa L.) solid waste as an antioxidant, urease and xanthine oxidase inhibitors. Food Chem 2017;235:119-26.
39. 39. Zolghadr L, ve ark. Molecular dynamics simulations, molecular docking, and kinetics study of kaempferol interaction on Jack bean urease: Comparison of extended solvation model. Food Sci Nutr 2022;10:3585-97.
40. 40. Saleem U, ve ark. Phytochemical analysis and wound healing studies on ethnomedicinally important plant Malva neglecta Wallr. J Ethnopharmacol 2020;249:112401.
41. 41. Hasimi N, ve ark. Chemical profile of Malva neglecta and Malvella sherardiana by LC-MS/MS, GC/MS and their anticholinesterase, antimicrobial and antioxidant properties with aflatoxin-contents. Marmara Pharm J 2017;21:471-84.
42. 42. Ramsay KST, ve ark. Chemical constituents of Stereospermum acuminatissimum and their urease and α-chymotrypsin inhibitions. Fitoterapia 2012;83:204-8.
43. 43. Güder A, Korkmaz H. Evaluation of in-vitro Antioxidant properties of hydroalcoholic solution extracts Urtica dioica L., Malva neglecta Wallr. and their mixture. Iranian J Pharm Res 2012;11(3):913-23.
44. 44. Dalar A, Türker M, Konczak I. Antioxidant capacity and phenolic constituents of Malva neglecta Wallr. and Plantago lanceolata L. from Eastern Anatolia Region of Turkey. J Herb Med 2012;2:42-51.
45. 45. Khan H, ve ark. Nutritional composition, antioxidant and antimicrobial activities of selected wild edible plants. J Food Biochem 2016;40:61-70.
46. 46. Hamayun R, Iqbal MS, Qadir MI. Assessment of analgesic, anti-inflammatory and anti-pyretic activities of Malva neglecta Wallr. in animal model. Bangladesh J Bot 2021;50(3):577-83.
47. 47. Akhtar MF, ve ark. Ameliorating effect of Malva neglecta Wallr on obesity and diabetes in Wistar rats: A mechanistic study. BioMed Res Int 2022;2022:2614599.
48. 48. Gürbüz I, ve ark. Anti-ulcerogenic activity of some plants used in folk medicine of Pinarbasi (Kayseri, Turkey). J Ethnopharmacol 2005;101(1-3):313-8.